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Naegleria fowleri: functional expression of the Nfa1 protein in transfected Naegleria gruberi by promoter modification.

Authors
Song, KJ; Jeong, SR; Park, S; Kim, K; Kwon, MH; Im, KI; Pak, JH; Shin, HJ
Citation
Experimental parasitology, 112(2):115-120, 2006
Journal Title
Experimental parasitology
ISSN
0014-48941090-2449
Abstract
To establish a transient transfection system in a Naegleria, we constructed three nfa1-pEGFP-N1 vectors by the promoter replacement and insertion of a nfa1 gene and transfected the DNAs into Naegleria gruberi using a lipid reagent. The transfection efficiency and usefulness of the three modified vectors were estimated by identifying the expressions of the EGFP and Nfa1 protein from N. gruberi. After transfection, the Nfa1 protein was functionally expressed on pseudopodia of N. gruberi. The strong GFP fluorescence was observed in N. gruberi transfected with the actin-nfa1-pEGFP-N1 vector, of which the CMV promoter region in the expression vector was replaced with the actin 5' UTR region. Additionally, when transgenic N. gruberi trophozoites were co-cultured with CHO target cells, the Nfa1 protein was also located on cytoplasm and pseudopodia, especially on a food cup that was formed in contact with target cells as it shown in pathogenic N. fowleri.
MeSH terms
Actins/geneticsAnimalsAntibodies, Protozoan/immunologyAntigens, Protozoan/biosynthesis*Antigens, Protozoan/geneticsAntigens, Protozoan/immunologyBase SequenceBlotting, WesternCHO CellsCricetinaeCricetulusGene Expression Regulation/physiology*Genetic VectorsGreen Fluorescent Proteins/geneticsMiceMice, Inbred BALB CMicroscopy, FluorescenceMolecular Sequence DataNaegleria/genetics*Naegleria/metabolismPlasmidsPromoter Regions, Genetic/genetics*Protozoan Proteins/biosynthesis*Protozoan Proteins/geneticsProtozoan Proteins/immunologyReverse Transcriptase Polymerase Chain ReactionTransfection
DOI
10.1016/j.exppara.2005.10.004
PMID
16321386
Appears in Collections:
Journal Papers > School of Medicine / Graduate School of Medicine > Microbiology
AJOU Authors
송, 경주정, 석률박, 선김, 경민권, 명희신, 호준
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